A Preliminary Experiment on Agrobacterium tumefaciens-Mediated Transformation of the P5CS1 GENE in Tall Fescue
محورهای موضوعی : مجله گیاهان زینتیMostafa K. Sarmast 1 , Hassan Salehi 2 , Mehrnaz Zarei 3
1 - Department of Horticultural Science, Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources (GUASNR), Gorgan 49138-43464, Golestan, Iran
2 - Department of Horticultural Science, College of Agriculture, Shiraz University, Shiraz, Iran
3 - Department of Horticultural Science, College of Agriculture, Shiraz University, Shiraz, Iran
کلید واژه: Drought stress, Gus assay, Uida gene, <, i>, F. arundinacea<, /i>, ,
چکیده مقاله :
Abiotic stress conditions can be imposed on plants through either poor water quality or adverse climate condition. These types of stress affect plant growth and development throughout their life cycle. Plants that are tolerant to drought are therefore valuable, and turf grasses have the potential to grow tolerant. Turf grasses provide ground cover and offer multiple benefits, from their aesthetic value which affects our daily lives to their preservation of land against erosion. The objectives of the present work were to find the ideal media for callus induction and regeneration followed by overexpressing the P5CS1 gene encoding proline for improving drought stress resistance in Festuca arundinacea Schreb. The husks of tall fescue seeds were removed and the seeds were then sliced longitudinally. The result of this treatment showed a greater callus induction efficiency in comparison with intact seeds in the culture media. In addition, an increase in regeneration efficiency was observed in media supplemented with 2,4-D along with BAP. The average of G418‒resistant calli obtained in the experiment was around 10%. The heterologous transformation of P5CS1 in F. arundinacea background was confirmed by PCR and the transient Gus assay. More than 90% of calli expressed the uida gene which can most probably convey resistance to drought stress.
تنشهای غیر زیستی ممکن است گیاهان را به دلیل شرایط محیطی نامناسب و یا کیفیت کم آب متاثر کنند. این تنشهای رشد و نمو گیاه را طی چرخههای رشدی تحت تاثیر قرار میدهند. به این دلیل گیاهان متحمل به خشکی ارزشمند بوده و سبز فرشها متحمل به خشکی میتوانند برای این منظور تولید شوند. این گونهها پوششی مناسبی روی سطح زمین ایجاد کرده و مزایایی مختلفی از جمله زیبایی و جلوگیری از فرسایش خاک دارند که زندگی روزانه ما را متاثر میکنند. هدف این پژوهش یافتن محیط کشت ایده ال برای القاء پینه و باززایی و به دنبال آن بیش بیان ژن P5CS1 کد کننده پرولین برای بهبود مقاومت به خشکی در چمانواش بلند بود. برای این منظور پوشش بذر حذف و سپس به صورت طولی بریده شد. این تیمار اثر قابل ملاحظهای در مقایسه با بذوری داشت که تحت این تیمار قرار نگرفته بودند. علاوه بر این راندمان باززایی در محیطهای دارای 2,4-D و بنزیل آدنین افزایش یافت. متوسط پینههای مقاوم به نشانگر گزینشگر G418 تنها 10% بود. حضور ژن P5CS1 در پینهها با استفاده از آزمون هیستوشیمیایی Gus و واکنش زنجیرهای پلیمراز با آغازگرهای اختصاصی تایید شد. بیش از 90% پینه ها قادر به بیان ژن uida بودند که این به احتمال و به طور غیر مستقیم بیان کننده القاء تحمل به خشکی در پینهها میباشد.
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